Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Usherin CRISPR/Cas9 KO Plasmid (m): sc-423630
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Usherin CRISPR/Cas9 Knockout (KO) Plasmid (m) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the Usherin genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Usherin Antibody (G-6): sc-515555
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Usherin CRISPR/Cas9 KO Plasmid (m) | sc-423630 | 20 µg | $397.00 |
Overview
Ush2a encodes usherin, a large extracellular matrix–associated transmembrane protein enriched at the periciliary membrane complex of sensory epithelia, where it supports mechanosensory structure and long-term cellular integrity. In mouse, usherin contributes to stereocilia organization and photoreceptor maintenance by coordinating protein complexes involved in cell adhesion, ciliary trafficking, and cytoskeletal coupling. Disruption of USH2A is strongly associated with Usher syndrome type II and retinitis pigmentosa, linking this gene to pathways governing cilium-dependent signaling and sensory neuron homeostasis. Accordingly, Ush2a loss-of-function models are used to interrogate ciliary transport, extracellular scaffold assembly, and degeneration-related stress responses in retina and inner ear contexts.
Usherin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ush2a gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ush2a together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the Ush2a open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish Usherin protein expression.
This CRISPR knockout system enables efficient generation of Ush2a-deficient cell models for investigation of Usherin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ush2a exon(s) critical for Usherin function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple Ush2a genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Usherin CRISPR/Cas9 KO Plasmid (m) and Usherin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ush2a locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Usherin HDR Plasmid (m) and Usherin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ush2a homology arms to support homology-directed repair at defined Ush2a target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.